Compact long-working-distance laser-diode-based photoacoustic microscopy with a reflective objective

Photoacoustic microscopy (PAM) has quickly developed into a noninvasive biomedical imaging technique to achieve detection, diagnosis, and monitoring. Compared with Q-switched neodymium-doped yttrium aluminum garnet or optical parametric oscillator lasers, a low-cost and small-size laser diode (LD) used as an alternative light source is conducive to achieving the miniaturization and integration for preclinical transformation. However, the LD's low peak output power needs the high numerical aperture objective to attain tight focus, which limits the working distance (WD) of the system in only 2-3 mm, resulting in not achieving the backward coaxial confocal mode. Here, we present a compact visible LD-based PAM system with a reflective objective to achieve a 22 mm long WD and a 10 mu m lateral resolution. Different depth subcutaneous microvascular networks in label-free mouse ears have successfully reappeared in vivo with a signal-to-noise ratio up to 14 dB by a confocal alignment. It will be a promising tool to develop into a handy tool for subcutaneous blood vessel imaging.